Related Product Information

Introduction

Isolate or deplete CD15+ cells directly from whole blood, buffy coat or MNC suspensions with Dynabeads CD15. For rapid and consistent results in protein or gene expression analysis, lyse the granulocytes while they are still attached to the beads and directly process for further molecular analysis.

Principle of Isolation

Dynabeads are mixed with the cell sample in a tube. The Dynabeads will bind to the target cells during a short incubation, and then the bead-bound cells are separated by a magnet.

Use a mixer that provides tilting and rotation of the tubes to ensure Dynabeads do not settle at the bottom of the tube.
When incubating Dynabeads and cells, the incubation temperature must be 2-8°C to reduce phagocytic activity and other metabolic processes.
Never use less than 25 μl (1 x 107) Dynabeads per ml of cell sample and at least 4 Dynabeads per target cell.

Table 1: Volume of Dynabeads added per ml of cell sample. The volumes can be scaled up as required.

Positive isolation

Depletion

Sample volume -

1 x 107 MNC*/ml

diluted whole blood

diluted buffy coat

1 ml

1 ml

Volume of Dynabeads

25 μl

50 μl

Total no. of cells
processed per product

2 x 109 cells

1 x 109 cells

* if the concentration of MNC is increased, the volume of Dynabeads must be increased accordingly. Cell concentration can be up to 1 x 108 cells per ml.

Depletion or Positive Isolation of CD15+ Cells

Add the appropriate volume of Dynabeads to the prepared sample according to table 1.

For positive isolation, discard the supernatant and wash the beadbound cells 3 times by resuspending in Buffer 1 to the original sample volume, and separate using a magnet. For molecular studies, lyse cells while still attached to the beads and transfer supernatant to a new tube for protein or gene expression analysis.

General Information

Invitrogen Dynal AS complies with the Quality System Standards ISO 9001:2000 and ISO 13485:2003.

Storage/Stability

This product is stable until the expiry date stated on the label when stored unopened at 2-8°C. Store opened vials at 2-8°C and avoid bacterial contamination. Keep Dynabeads in liquid suspension during storage and all handling steps, as drying will result in reduced performance. Resuspend well before use.

Warnings And Limitations

This product is for research use only. Not intended for any animal or human therapeutic or diagnostic use unless otherwise stated. Follow appropriate laboratory guidelines. This product contains 0.02% sodium azide as a preservative, which is cytotoxic.

Avoid pipetting by mouth!

Sodium azide may react with lead and copper plumbing to form highly explosive metal azides. When disposing through plumbing drains, flush with large volumes of water to prevent azide build up. Certificate of Analysis (CoA) is available upon request. Material Safety Data Sheet (MSDS) is available at .